Unattached kinetochores rather than intrakinetochore tension arrest mitosis in taxol-treated cells

The Journal of Cell Biology
Valentin MagidsonA Khodjakov

Abstract

Kinetochores attach chromosomes to the spindle microtubules and signal the spindle assembly checkpoint to delay mitotic exit until all chromosomes are attached. Light microscopy approaches aimed to indirectly determine distances between various proteins within the kinetochore (termed Delta) suggest that kinetochores become stretched by spindle forces and compact elastically when the force is suppressed. Low Delta is believed to arrest mitotic progression in taxol-treated cells. However, the structural basis of Delta remains unknown. By integrating same-kinetochore light microscopy and electron microscopy, we demonstrate that the value of Delta is affected by the variability in the shape and size of outer kinetochore domains. The outer kinetochore compacts when spindle forces are maximal during metaphase. When the forces are weakened by taxol treatment, the outer kinetochore expands radially and some kinetochores completely lose microtubule attachment, a condition known to arrest mitotic progression. These observations offer an alternative interpretation of intrakinetochore tension and question whether Delta plays a direct role in the control of mitotic progression.

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Methods Mentioned

BETA
light microscopy
FCS
transfection
transfections

Software Mentioned

Adobe Illustrator
SoftWoRx
Illustrator
Windows
lsqcurvefit
ImageJ
MATLAB
Adobe Photoshop
IMOD
SHREC

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